Backbone dynamics of Tet repressor alpha8intersectionalpha9 loop.

B Vergani; M Kintrup; W Hillen; H Lami; E Piémont; E Bombarda; P Alberti; S M Doglia; M Chabbert

doi:10.1021/bi9912591

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Backbone dynamics of Tet repressor alpha8intersectionalpha9 loop.

B Vergani·M Kintrup·W Hillen·H Lami·E Piémont·E Bombarda·P Alberti·S M Doglia·M Chabbert

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Abstract

A set of single Trp mutants of class B Tet repressor (TetR), in which Trp residues are located from positions 159 to 167, has been engineered to investigate the dynamics of the loop joining the alpha-helices 8 and 9. The fluorescence anisotropy decay of most mutants can be described by the sum of three exponential components. The longest rotational correlation time, 30 ns at 10 degrees C, corresponds to the overall rotation of the protein. The shortest two components, on the subnanosecond and nanosecond time scale, are related to internal motions of the protein. The initial anisotropy, in the 0.16-0.22 range, indicates the existence of an additional ultrafast motion on the picosecond time scale. Examination of physical models for underlying motions indicates that librational motions of the Trp side chain within the rotameric chi(1) x chi(2) potential wells contribute to the picosecond depolarization process, whereas the subnanosecond and nanosecond depolarization processes are related to backbone dynamics. In the absence of inducer, the order parameters of these motions, about 0.90 and 0.80 for most positions, indicate limited flexibility of the loop backbone. Anhydrotetracycline binding to TetR induces an increased mobility of the loop on the nanosecond time scale. This suggests that entropic factors might play a role in the mechanism of allosteric transition.

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Bacterial Proteins Energy Transfer Fluorescence Polarization Peptide Fragments Protein Conformation Protein Structure, Secondary Repressor Proteins Spectrometry, Fluorescence Tetracycline Tetracyclines Thermodynamics Tryptophan Journal Article Research Support, Non-U.S. Gov't

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B Vergani, M Kintrup, W Hillen, H Lami, E Piémont, E Bombarda, P Alberti, S M Doglia, M Chabbert. 2000-03-14. Backbone dynamics of Tet repressor alpha8intersectionalpha9 loop.. https://doi.org/10.1021/bi9912591

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